16 SrRNA基因聚合酶链反应检测细菌感染的研究

Polymerase Chain Reaction Amplification of 16 SrRNA Gene for the Detection of Bacterial Infection

目的：探索快速可靠的检测细菌感染的新方法。方法：通过自行设计合成的细菌１６ＳｒＲＮＡ基因高度保守区引物，对２０ 种标准菌株、１２ 种２７ 株临床分离的细菌株、人基因组ＤＮＡ及巨细胞病毒进行聚合酶链反应（ＰＣＲ）扩增。结果：对所测细菌株均获得３７１ ｂｐ 扩增产物，而与人基因组ＤＮＡ、巨细胞病毒无交叉阳性反应，ＰＣＲ最低能检测ｌｐｇ 大肠杆菌ＤＮＡ。结论：建立了用共同引物ＰＣＲ扩增以判断是否存在细菌感染的方法，该方法检测快速，敏感性和特异性高。

Objective:To study a rapid and reliable method for the detection of bacteria infection.Methods:Twenty standard bacterial strains,27 clinical bacterial isolates of 12 species,total human DNA,and cytomegalovirus were studied with polymerase chain reaction (PCR) amplification using a primer pair of 16 SrRNA gene.Results:The 16 SrRNA gene primers successfully amplified DNA from the standard strains and patient's isolated bacterial strains,PCR products were 371 bp in length,but human DNA and cytomegalovirus showed no amplification products.The sensitivity of detection of amplification showed that it was possible to detect reproducibly a band with template amounts of E.coli DNA as low as 1 pg.Conclusions:PCR amplification of bacterial DNA using highly conserved sequences for the detection of bacteria is feasible,and it offers advantages of speed,sensitivity and specificity.